Transport engineering for improving the production and secretion of valuable alkaloids in Escherichia coli
Microorganisms can be metabolically engineered to produce specialized plant metabolites. However, these methods are limited by low productivity and intracellular accumulation of metabolites. We sought to use transport engineering for producing reticuline, an important intermediate in the alkaloid bi...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2021-12-01
|
| Series: | Metabolic Engineering Communications |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2214030121000249 |
| _version_ | 1819177489718575104 |
|---|---|
| author | Yasuyuki Yamada Miya Urui Hidehiro Oki Kai Inoue Haruyuki Matsui Yoshito Ikeda Akira Nakagawa Fumihiko Sato Hiromichi Minami Nobukazu Shitan |
| author_facet | Yasuyuki Yamada Miya Urui Hidehiro Oki Kai Inoue Haruyuki Matsui Yoshito Ikeda Akira Nakagawa Fumihiko Sato Hiromichi Minami Nobukazu Shitan |
| author_sort | Yasuyuki Yamada |
| collection | DOAJ |
| description | Microorganisms can be metabolically engineered to produce specialized plant metabolites. However, these methods are limited by low productivity and intracellular accumulation of metabolites. We sought to use transport engineering for producing reticuline, an important intermediate in the alkaloid biosynthetic pathway. In this study, we established a reticuline-producing Escherichia coli strain into which the multidrug and toxic compound extrusion transporter Arabidopsis AtDTX1 was introduced. AtDTX1 was selected due to its suitable expression in E. coli and its reticuline-transport activity. Expression of AtDTX1 enhanced reticuline production by 11-fold, and the produced reticuline was secreted into the medium. AtDTX1 expression also conferred high plasmid stability and resulted in upregulation or downregulation of several genes associated with biological processes, including metabolic pathways for reticuline biosynthesis, leading to the production and secretion of high levels of reticuline. The successful employment of a transporter for alkaloid production suggests that the proposed transport engineering approach may improve the biosynthesis of specialized metabolites via metabolic engineering. |
| first_indexed | 2024-12-22T21:27:28Z |
| format | Article |
| id | doaj.art-6aebad267daf4f2783262f7cdf063c8a |
| institution | Directory Open Access Journal |
| issn | 2214-0301 |
| language | English |
| last_indexed | 2024-12-22T21:27:28Z |
| publishDate | 2021-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Metabolic Engineering Communications |
| spelling | doaj.art-6aebad267daf4f2783262f7cdf063c8a2022-12-21T18:12:00ZengElsevierMetabolic Engineering Communications2214-03012021-12-0113e00184Transport engineering for improving the production and secretion of valuable alkaloids in Escherichia coliYasuyuki Yamada0Miya Urui1Hidehiro Oki2Kai Inoue3Haruyuki Matsui4Yoshito Ikeda5Akira Nakagawa6Fumihiko Sato7Hiromichi Minami8Nobukazu Shitan9Laboratory of Medicinal Cell Biology, Kobe Pharmaceutical University, Motoyamakita-machi, Higashinada-ku, Kobe, 658-8558, JapanLaboratory of Medicinal Cell Biology, Kobe Pharmaceutical University, Motoyamakita-machi, Higashinada-ku, Kobe, 658-8558, JapanLaboratory of Medicinal Cell Biology, Kobe Pharmaceutical University, Motoyamakita-machi, Higashinada-ku, Kobe, 658-8558, JapanLaboratory of Medicinal Cell Biology, Kobe Pharmaceutical University, Motoyamakita-machi, Higashinada-ku, Kobe, 658-8558, JapanLaboratory of Medicinal Cell Biology, Kobe Pharmaceutical University, Motoyamakita-machi, Higashinada-ku, Kobe, 658-8558, JapanLaboratory of Medicinal Cell Biology, Kobe Pharmaceutical University, Motoyamakita-machi, Higashinada-ku, Kobe, 658-8558, JapanResearch Institute for Bioresources and Biotechnology, Ishikawa Prefectural University, Nonoichi-machi, Ishikawa, 921-8836, JapanDepartment of Plant Gene and Totipotency, Division of Integrated Life Science, Graduate School of Biostudies, Kyoto University, Kyoto, 606-8502, Japan; Graduate School of Science, Osaka Prefecture University, Sakai, 599-8531, JapanResearch Institute for Bioresources and Biotechnology, Ishikawa Prefectural University, Nonoichi-machi, Ishikawa, 921-8836, JapanLaboratory of Medicinal Cell Biology, Kobe Pharmaceutical University, Motoyamakita-machi, Higashinada-ku, Kobe, 658-8558, Japan; Corresponding author.Microorganisms can be metabolically engineered to produce specialized plant metabolites. However, these methods are limited by low productivity and intracellular accumulation of metabolites. We sought to use transport engineering for producing reticuline, an important intermediate in the alkaloid biosynthetic pathway. In this study, we established a reticuline-producing Escherichia coli strain into which the multidrug and toxic compound extrusion transporter Arabidopsis AtDTX1 was introduced. AtDTX1 was selected due to its suitable expression in E. coli and its reticuline-transport activity. Expression of AtDTX1 enhanced reticuline production by 11-fold, and the produced reticuline was secreted into the medium. AtDTX1 expression also conferred high plasmid stability and resulted in upregulation or downregulation of several genes associated with biological processes, including metabolic pathways for reticuline biosynthesis, leading to the production and secretion of high levels of reticuline. The successful employment of a transporter for alkaloid production suggests that the proposed transport engineering approach may improve the biosynthesis of specialized metabolites via metabolic engineering.http://www.sciencedirect.com/science/article/pii/S2214030121000249AlkaloidsMetabolic engineeringTransport engineeringE. coliReticulineAtDTX1 |
| spellingShingle | Yasuyuki Yamada Miya Urui Hidehiro Oki Kai Inoue Haruyuki Matsui Yoshito Ikeda Akira Nakagawa Fumihiko Sato Hiromichi Minami Nobukazu Shitan Transport engineering for improving the production and secretion of valuable alkaloids in Escherichia coli Metabolic Engineering Communications Alkaloids Metabolic engineering Transport engineering E. coli Reticuline AtDTX1 |
| title | Transport engineering for improving the production and secretion of valuable alkaloids in Escherichia coli |
| title_full | Transport engineering for improving the production and secretion of valuable alkaloids in Escherichia coli |
| title_fullStr | Transport engineering for improving the production and secretion of valuable alkaloids in Escherichia coli |
| title_full_unstemmed | Transport engineering for improving the production and secretion of valuable alkaloids in Escherichia coli |
| title_short | Transport engineering for improving the production and secretion of valuable alkaloids in Escherichia coli |
| title_sort | transport engineering for improving the production and secretion of valuable alkaloids in escherichia coli |
| topic | Alkaloids Metabolic engineering Transport engineering E. coli Reticuline AtDTX1 |
| url | http://www.sciencedirect.com/science/article/pii/S2214030121000249 |
| work_keys_str_mv | AT yasuyukiyamada transportengineeringforimprovingtheproductionandsecretionofvaluablealkaloidsinescherichiacoli AT miyaurui transportengineeringforimprovingtheproductionandsecretionofvaluablealkaloidsinescherichiacoli AT hidehirooki transportengineeringforimprovingtheproductionandsecretionofvaluablealkaloidsinescherichiacoli AT kaiinoue transportengineeringforimprovingtheproductionandsecretionofvaluablealkaloidsinescherichiacoli AT haruyukimatsui transportengineeringforimprovingtheproductionandsecretionofvaluablealkaloidsinescherichiacoli AT yoshitoikeda transportengineeringforimprovingtheproductionandsecretionofvaluablealkaloidsinescherichiacoli AT akiranakagawa transportengineeringforimprovingtheproductionandsecretionofvaluablealkaloidsinescherichiacoli AT fumihikosato transportengineeringforimprovingtheproductionandsecretionofvaluablealkaloidsinescherichiacoli AT hiromichiminami transportengineeringforimprovingtheproductionandsecretionofvaluablealkaloidsinescherichiacoli AT nobukazushitan transportengineeringforimprovingtheproductionandsecretionofvaluablealkaloidsinescherichiacoli |